Molecules 2011, 16, 5875-5885; doi:10.

3390/molecules16075875

OPEN ACCESS

molecules
ISSN 1420-3049
www.mdpi.com/journal/molecules
Article

Comparative Properties of Amazonian Oils Obtained by

Different Extraction Methods
Bianca Silva Ferreira 1, Camila Guimarães de Almeida 1, Lara Pereira Faza 1,
Angelina de Almeida 1, Cláudio Galuppo Diniz 2, Vânia Lúcia da Silva 2,
Richard Michael Grazul 1 and Mireille Le Hyaric 1,*
1
Departamento de Química, Instituto de Ciências Exatas, Universidade Federal de Juiz de Fora,
Rua José Lourenço Kelmer, s/n – Campus Universitário, Bairro Martelos, 36036-900,
Juiz de Fora-MG, Brazil
2
Departamento de Parasitologia, Microbiologia e Imunologia, Instituto e Ciências Biológicas,
Universidade Federal de Juiz de Fora, 36036-900, Juiz de Fora, Brazil

* Author to whom correspondence should be addressed; E-Mail: mireille.hyaric@ufjf.edu.br;

Tel.: +55-32-2102-3310; Fax: +55-32-2102-3310.

Received: 23 June 2011; in revised form: 30 June 2011 / Accepted: 1 July 2011 /
Published: 13 July 2011

Abstract: Pequi (Caryocar brasiliense Camb.), babaçu (Orbignya phalerata Mart.), buriti
(Mauritia flexuosa), and passion fruit (Passiflora edulis) oils were studied to determine
their antibacterial, antioxidant and cytotoxic activities, as well as their total phenol and
carotenoid contents. The fatty acid contents were determined by GC-MS. The three types
of passion fruit oils studied were refined, cold pressed or extracted from seeds in a Soxhlet
apparatus. The oils thus obtained showed differences in antioxidant activity and carotenoid
content, but were similar in regard to total phenols. Buriti and pequi had the highest
carotenoid contents, while refined and cold pressed passion fruit oil displayed the highest
antioxidant activity. Pequi oil was the only oil to display antibacterial and
cytotoxic activity.

Keywords: antibacterial; antioxidant; cytotoxicity; Amazonian oils; pequi; passion fruit

Molecules 2011, 16 5876

1. Introduction

The Amazon is the world's most important ecosystem, which includes territories belonging to nine
South American countries, with 61% being in Brazil. Of the estimated 30 million species of plants
found in the Amazon, only a few have been studied and identified so far, based on popular knowledge
and scientific studies. Turning these resources into renewable raw materials for industry is a major
challenge [1]. Babaçu (Orbignya phalerata Mart.), buriti (Mauritia flexuosa), and passion fruit
(Passiflora edulis) are examples of economically important species, whose oils and fats have found
various applications in the food, pharmaceutical, cosmetic, and textile industries. As the use of
Amazonian vegetable oils has increased in recent years, it is important to aquire a better knowledge of
their properties in order to optimize the use of these materials. In this work we studied the
antibacterial, antioxidant and cytotoxic activities of pequi (Caryocar brasiliense Camb.), babaçu, buriti
and passion fruit seed oils. We also determined the β-carotene content and the total phenol contents of
each oil. Three types of passion fruit oil were used: industrial refined oil, cold pressed oil and the oil
obtained in the laboratory by Soxhlet extraction of the seeds.
The edible oil and fruits of pequi are used in cooking as well as in traditional medicine for the
treatment of colds, coughs, bronchitis, edema and burns [2]. The oil extracted from the nut has
applications in soaps and skin emulsions. Babaçu is one of the most important palm trees in Brazil. All
the parts of the plants are used, providing food, fuel, fiber, construction materials and more [3]. The
edible oil represents 65% of the seed and is used in the fabrication of soaps, surfactants and margarine.
Buriti oil is extracted from the fruit of a palm tree native to Brazil. The edible oil, used in cooking, is
rich in monounsaturated fatty acids, [4] and natural antioxidants [5]. Buriti oil has found applications
in the cosmetic industry due to its emollient properties and can be used as an adjuvant in sun
protection [6]. Roots and aerial parts of Passiflora species are traditionally used in many countries for
its anxiolytic, sedative, diuretic, analgesic, antimicrobial and many other biological effects [7]. Fruits,
leaves and seed extracts also display cytotoxic and antioxidant activity, due to the presence of
carotenoids, polyphenols and tocopherols [8]. The seed oil, a powerful moisturizing agent, is currently
used in many cosmetic formulations.

2. Results and Discussion

2.1. Physicochemical Analysis

Usual physicochemical parameters of the oils such as density, saponification value (SV), iodine
value (IV), peroxide value (PV), acid values (AV), and ester value (EV) were determined (Table 1).
The values obtained were consistent with the manufacturer’s analytical reports for the refined oils and
with values found in the literature.
The high peroxide (28.63) and acid values (10.86) found for buriti oil indicate that it should not be
used for human consumption, as the recommendation for edible oils is a peroxide value less than
10 meq O2/kg [14]. When purchased, this oil was stored in a reused polyethylene terephthalate bottle
exposed to light, with no indication of the date of manufacture: the prolonged improper handling may
have led to lipid peroxidation and partial hydrolysis of the oil.
Molecules 2011, 16 5877

Table1. Physicochemical analysis.

Density SV IV PV AV (% Oleic) EV (%)

Pequi [9,10] 0.967 ± 0.002 206.8 ± 5.9 50.0 ± 0.93 7.94 ± 0.01 5.4 ± 0.03 97.38 ± 5.9
Babaçu [11] 0.920 ± 0.002 236.9 ± 2.7 18.3 ± 0.50 nd 1.4 ± 0.001 99.41 ± 2.7
Buriti [12] 0.909 ± 0.004 202.4 ± 1.8 61.8 ± 0.49 28.63 ± 0.04 10.9 ± 0.04 87.40 ± 1.9
Passion fruit [13] 0.920 ± 0.002 173.4 ± 2.2 236.8 ± 0.46 3.69 ± 0.17 0.2 ± 0.04 99.87 ± 2.3
(Refined)
Passion fruit 0.922 ± 0.003 194.3 ± 0.0 109.0 ± 0.52 nd 0.7 ± 0.02 99.62 ± 0.0
(Cold pressed)
Passion fruit 0.946 ± 0.002 170.2 ± 2.7 120.3 ± 2,31 nd 1.5 ± 0.03 99.14 ± 2.7
(Soxhlet
extraction)
nd: no peroxide was detected.

2.2. Fatty Acid Profile

2.2.1. GC-MS

GC-MS analysis was used to estimate the content of the principal fatty acids (unsaturated fatty acids
and total saturated fatty acids) of the oils examined in this work. The calculated values were in
agreement with the literature data (Table 2). Oleic acid (C18:1) and palmitic acid (C16:0) were the
major fatty acids present in pequi and buriti oils. Babaçu oil was the most saturated, with a high
content of lauric acid (C12:0; 54.7%). The three passion fruit oils were rich in linoleic acid (C18:2),
and contained large amounts of palmitic acid and oleic acid.

Table 2. Fatty acids composition (mean percentage) of Amazonian vegetable oils.

Passion Passion Passion
Pequi [10] Babaçú [11] Buriti [12]
fruit [15] fruit [15] fruit c [16]
a b

C6:0 nd 3.3 nd nd nd nd
C8:0 nd 9.2 nd nd nd nd
C10:0 nd 9.6 nd nd nd nd
C12:0 nd 54.7 nd nd nd nd
C14:0 nd 11.8 nd nd nd nd
C16:0 34.5 4.8 16.6 21.6 14.1 12.8
C18:1 65.5 6.5 83.4 21.9 13.2 10.7
C18:2 nd nd nd 56.4 72.6 72.6
a b c
Refined oil; Cold pressed oil; Soxhlet extracted oil; nd = not detected.
Molecules 2011, 16 5878

2.3. Antibacterial Screening

Representative Gram-positive and Gram-negative bacteria were used for the evaluation of
antimicrobial activity. The activity was detected by observing and measuring the diameter of microbial
growth inhibition zones surrounding the different oils which were added to the test system. The results
presented in Table 3 show that only pequi oil displayed antibacterial activity against one of the bacteria
(P. aeruginosa). The results suggest at least a bacteriostatic activity of pequi oil and its potential use as
a preservative. Recent literature reports that oil obtained from Caryocar coriaceum, another species of
the generus Caryocar, shows potential antibacterial activity against P. aeruginosa, S. aureus and
S. cholerasius at concentrations of 1.25% [17]. The other oils evaluated in this study did not show any
antibacterial activity, being harmless against all the tested bacteria.

2.4. Antioxidant Activity

2.4.1. Qualitative Assay

All the oils bleached the DPPH solution (0.2% in MeOH) purple color in the TLC autography
qualitative assay, showing that all of them are potential antioxidants.

Table 3. Antibacterial activity.

Inhibition zone (mm)
S. epidermidis S. aureus P. aeruginosa E. coli
Pequi - - 7 -
Babaçu - - - -
Buriti - - - -
Passion fruit (Refined) - - - -
Passion fruit (Cold pressed) - - - -
Passion fruit (Extracted) - - - -
DMSO - - - -

2.4.2. Quantitative Analysis

Quantitative assay revealed varying degrees of antioxidant activity of the oils. Refined and cold
pressed passion fruit oil showed the best antioxidant capacity (Table 4). The Soxhlet extract was half
as active, probably due to the loss of natural antioxidant compounds caused by heating. Pequi oil was
expected to display a higher antioxidant activity, as the pulp contains gallic and quinic acids, powerful
antioxidants [18]. Buriti oil, rich in tocopherols and carotenoids13 showed good activity, comparable
to refined passion fruit oil.
Molecules 2011, 16 5879

Table 4. Antioxidant activity.

Oil EC50 (mg/mL)
Pequi 15.54 ± 2.1
Babaçu 70.57 ± 0.4
Buriti 7.70 ± 0.6
Passion fruit (Refined) 5.74 ± 0.1
Passion fruit (Cold pressed) 7.12 ± 0.2
Passion fruit (Soxhlet) 16.84 ± 0,5
Ácido ascórbico (μg/mL) 0.04 ± 0,0

2.5. Carotene Determination and Total Phenol Content

The colored buriti and pequi oil are known to contain β-carotene as the main carotenoid in their
compositions [19,20]. Pequi, buriti and passion fruit oils were the richest in carotenoids (Table 5),
which can be related to their high antioxidant activity. As expected, babaçu oil did not present a high
content of carotenoids. Considering the antioxidant activity of the passion fruit oils we were expecting
a much higher content of carotenoids in the refined oil. It appeared that it contains less carotenoids
than the Soxhlet extract, which also has the highest antioxidant activity. This higher carotene content
can be explained by the solvent extraction with an apolar solvent.

Table 5. Total carotene content.

Oil Carotene content μg/g
Pequi 274.9 ± 3.4
Babaçu 19.8 ± 0.5
Buriti 692.9 ± 6.8
Passion fruit (Refined) 2.3 ± 0.15
Passion fruit (Cold pressed) 4.6 ± 0.1
Passion fruit (Soxhlet) 19.7 ± 0.3

As phenols are also responsible for the antioxidant activity of many other classes as of plants and
their extracts [21], we also determined the total phenol contents of each oil (Table 6). The Soxhlet
extracted passiflora oil has a lower phenol content than the analogous pressed oil: it is well known that
polyphenols are destroyed by heat [22]. The results show that there is not a direct correlation between
phenol contents and antioxidant activities. The total phenol content was higher for passion fruit oil
which displayed a lower antioxidant activity. Pequi and babaçu oils had the lowest phenol contents, but
the highest antioxidant activity.
Molecules 2011, 16 5880

Table 6. Total phenol content.

Oil Total phenol content
(g/g)
Pequi 229.1 ± 1.65
Babaçu 288.0 ± 1.5
Buriti 309.9 ± 2.9
Passion fruit (Refined) 350.4 ± 1.5
Passion fruit (Cold pressed) 349.6 ± 2.9
Passion fruit (Soxhlet) 339.6 ± 2.8

2.5.1. Cytotoxic Activity

The brine shrimp lethality assay was employed as a toxicity screen. The lethal dose (LD50) is the
amount of a substance which causes the death of 50% of a group of test organisms. LD50 values lower
than 1,000 mg/mL indicate toxicity. From the results (Table 7) the tested pequi oil was the only toxic
one, suggesting that it should be carefully used. Previous reports have described biological activity and
toxicity of leaf extracts of Caryocar brasiliense [23,24], but to our knowledge, the toxicity of the oil
obtained from the fruit has not been reported yet. Further studies should be performed to determine if
the toxicity is specific to the specific sample studied or if it is common to pequi oils.

Table 7. Cytotoxic activity.

Oil LD50 (μg/mL)
Buriti >1000
Pequi 827.62 ± 4,67
Babaçu >1000
Passion fruit (Refined) >1000
Passion fruit (Cold pressed) >1000
Passion fruit (Soxhlet) >1000

3. Materials and Methods

3.1. Vegetable Oils

Artisanal pequi, babaçu and buriti oils were purchased from the same producer in a popular market
in Januária, Minas Gerais, Brazil. These oils were obtained by cooking the pulp in boiling water,
separating the supernatant oil, then drying the oil in a pan until it lost its opacity, and finally filtering.
Refined passion fruit oil was purchased from Emfal Empresa Fornecedora de Álcool Ltda, Betim,
Minas Gerais, Brazil. Cold pressed passion fruit oil was generously furnished by Extrair, Neale & Reis
Indústria e Comércio de Óleos Naturais Ltda, Bom Jesus do Itabapoana – RJ, Brazil.
Molecules 2011, 16 5881

For the Soxhlet extraction we used seeds from fruits purchased in local markets. The seeds were
separated from the pulp in a kitchen food processor, washed with water and frozen until used in order
to avoid any modification due to exposure to heat or light. After thawing, the seeds were dried at 40 °C
for 2 hours and ground before extraction in a Soxhlet apparatus with petroleum ether for 4 hours. After
evaporation of the solvent, a yellow, limpid oil was obtained (yield 13%). The oil was frozen until it
was used.

3.2. Physicochemical Analysis

The following physicochemical properties of oils were determined: density, saponification value
(SV), iodine value (IV), peroxide value (PV), free acid (FA) content, according to AOCS official
methods [25]. All analyses were performed in triplicate.

3.3. Fatty Acid Profile

The fatty acid profile was determined as fatty acid methyl esters by gas chromatography-mass
spectroscopy. The methyl esters were prepared using the AOCS method (AOCS Ce 2-66) [26].
Separation of fatty acid esters was performed on a Shimadzu GC-2010 Gas Chromatograph equipped
with a Restek RTX-2330 capillary column (60 m × 0.25 mm × 0.2 mm). The column temperature was
programmed at 130 °C for 10 min, then increased to 230 °C at 5 °C/min with a final isothermal period
of 13 min. Hydrogen was used as carrier gas with constant linear velocity of 25 cm/sec−1. The injector
temperature was set at 250 °C, with a split ratio of 1:10. The flame ionization detector temperature was
250 °C. Fatty acid methyl esters (FAMEs) were identified by comparison of retention times with
authentic standards (Supelco 37 comp. FAME mix 10 mg/mL in CH2Cl2), and quantification was
performed by the internal normalization method.

3.4. Antibacterial Screening

The antibacterial assay was performed using the solid medium diffusion technique [27]. Briefly,
from an overnight culture in Tryptic Soy Agar (Himedia, Mumbai, India), 0.5 McFarland bacterial
suspensions were obtained and 0.1 mL of each suspension (approximately 108 CFU/mL) was spread on
a sterile Mueller-Hinton Agar plate (Himedia, Mumbai, India). After a period of 5-10 min, wells of
5 mm diameter were made in the inoculated Mueller-Hinton plates. Each well was filled with 0.1 mL
of a DMSO solution of each tested oil (10 mg/mL). The plates were then aerobically incubated at
35.5 °C for 24 h. The antibacterial activity was determined by measuring the growth inhibition zones
recorded in mm in each plate. Nitrofurazone was used as a positive control and DMSO was used as the
negative control. The tests were performed in duplicate in which the oils were tested against two Gram
positive bacteria strains (Staphylococcus epidermidis ATCC12228 and Staphylococcus aureus
ATCC25923) and two Gram negative bacteria strains (Escherichia coli ATTC11229 and Pseudomonas
aeruginosa ATTC27853).
Molecules 2011, 16 5882

3.5. Antioxidant Activity

3.5.1. Qualitative DPPH Assay

Samples of the different oils were dissolved in dichloromethane and spotted on a thin layer
chromatography plate. After total evaporation of the solvent, the plates were sprayed with
2,2-diphenyl-2-picryl-hydrazine (DPPH, Sigma Aldrich, 0.2 mM in methanol). The antioxidant activity
was detected after 30 min by the presence of yellow or white spots arising from the reduction of
DPPH, against a purple background. β-carotene (0.2 g/L in methanol) was used as positive standard.

3.5.2. Quantitative Analysis

The antioxidant activity of tested oils was determined using the spectroscopic DPPH free radical
scavenging assay [28]. Five concentrations (100, 150, 200, 250 and 300 mg/mL in chloroform) were
prepared for each oil, then 0.1 mL of the different mixtures was added to 3.9 mL of a freshly prepared
solution of DPPH (0.06 mM in methanol). The absorbance (OD) was measured at 515 nm after
20 min. Ascorbic acid was used as a positive control, DPPH solution without oil solution was used as
control and methanol was used as blank. The measurements were performed in triplicate. The
antioxidant activity was obtained from the following equation:
Antioxidant activity (%) = [(OD DPPH − OD oil) / (OD DPPH)] × 100
Percent antioxidant activities were then plotted against concentration. The antioxidant activity of
the oils was expressed as EC50, defined as the concentration in mg/mL required to scavenge 50% of the
DPPH free radical.

3.6. Carotene Determination

The quantification was performed by the external standard method, using β-carotene
(Sigma-Aldrich) as a reference. The absorbance of the solutions was measured at 436 nm. A solution
of β-carotene (0.2 g/L) was prepared in dichloromethane and diluted to obtain final concentrations of
1.26, 2.52, 3.78, 5.04 and 6.30 mg/L which were used for the construction of the standard curve. The
concentration of carotenoids in the extract was calculated from the equation straight line, obtained by
linear regression.

3.7. Total Phenol Content

The total phenol contents in the oils were determined using the Folin-Ciocalteau reagent and gallic
acid as standard [29]. The absorbance of the solutions was measured at 765 nm. A solution of gallic acid
was prepared by dissolving dry gallic acid (0.5 g) in ethanol (10 mL) and then diluting to 100 mL with
water and diluted to obtain phenol concentrations of 0, 50, 100, 150, 250 and 500 mg/L which were used
for the construction of the calibration curve. The sample (20 mL) was added to water (1.58 mL),
Folin-Ciocalteau reagent (100 mL) and sodium carbonate solution (100 g/L, 300 mL). After 30 min of
reaction at room temperature, the absorbance was measured at 765 nm in a Shimadzu 160-UV
spectrophotometer. Tests were carried out in triplicate.
Molecules 2011, 16 5883

3.8. Cytotoxicity Screening

The cytoxicity test was performed on the Artemia nauplii (brine shrimp larvae) using the Meyer
method [30]. The eggs were acquired from an aquarium shop (São Paulo, Brazil) and hatched in
artificial sea water, prepared by dissolving NaCl (69 g), MgCl2·6H2O (59.3 g), Na2SO4 (132 g), CaCl2
(6.3 g) and KCl (2.1 g) in distilled water (3 L). After homogenization of the sea water, the pH was
adjusted to between 8 and 9 using a Na2CO3 solution (0.1 M). Brine shrimp eggs were incubated in the
artificial seawater at room temperature for 48 hours. An aquarium with a partition was used to allow
the migration of the larvae to the lit side. The larvae were fed with biological yeast (18 mg/L in
artificial sea water).
The oils were first dissolved in DMSO/Tween (1:1) and the resulting solutions were diluted to
obtain the following concentrations: 50 mg/mL, 25 mg/mL, 5 mg/mL, 0.5 mg/mL and 0.05 mg/mL.
Aliquots of 0.1 mL of each dilution were added to test tubes containing artificial seawater (4.9 mL)
and 10 nauplii. The negative control was carried out in DMSO/Tween (1%) in seawater, and the
positive control with K2Cr2O7 (200 μg/mL). Nauplii still alive after 24 h were counted and the lethal
concentration (LC50) was calculated plotting the percentage mortality versus concentration. All tests
were performed in triplicate.

3.9. Statistical Analysis

The results presented in this study correspond to the mean of three replicates ± standard deviation.
If the variance was homogeneous, the data were assessed by one-way analysis of variance (ANOVA)
followed by the Tukey-Kramer test. All calculations were performed using the program Microcal
Origin 6.0.

4. Conclusions

Among the six Amazonian oils studied in this work, only pequi oil displayed antibacterial and
cytotoxic activity. The three passion fruit oils differed in the way they were obtained and also differed
in their properties: the refined oil displayed a higher antioxidant activity, but contained fewer
carotenoids than the cold pressed oil. The high peroxide and acid contents found for buriti oil points to
the lack of quality control in the sail of artisanal products, as this particular oil should not have been
commercialized.

Conflicts of Interest

The authors declare that there are no conflicts of interest.

Acknowledgements

The authors gratefully acknowledge FAPEMIG, CAPES and CNPq for fellowships and financial
support and Programa de Apoio à Publicação/Pró-reitoria de Pesquisa/Universidade Federal de Juiz de
Fora (PROPESQ/UFJF).
Molecules 2011, 16 5884

References

1 Gilbert, B. Economic Plants of the Amazon. In Chemistry of the Amazon: Biodiversity, Natural
Products and Environmental Issues; Seidl, P., Gottlieb, O.R., Kaplan, M.A., Eds.; ACS
Symposium Series: Washington, DC, USA, 1995; pp. 9-33.
2 Agra, M.F.; Freitas, P.F.; Barbosa-Filho, J.M. Synopsis of the plants known as medicinal and
poisonous in Northeast of Brazil. Rev. Bras. Farmacog. 2007, 17, 17114-17140.
3 Balick, M.J. The use of palms by the Apinayé and Guajajara Indians of northeastern Brazil. Adv.
Econ. Bot. 1988, 6, 65-90.
4 Albuquerque, M.L.S.; Guedes, I.; Alcantara, J.R.P.; Moreira, S.G.C.; Barbosa Neto, N.M.;
Correa, D.S.; Zilio, S.C. Characterization of buriti (Mauritia flexuosa L.) oil by absorption and
emission spectroscopies. J. Braz. Chem. Soc. 2005, 16, 1113-1117.
5 Franc, L.F.; Reber, G.; Meireles, M.A.A.; Machado, N.T.; Brunner, G. Supercritical extraction of
carotenoids and lipids from buriti (Mauritia flexuosa), a fruit from the Amazon region.
J. Supercritical Fluids 1999, 14, 247-256.
6 Zanatta, C.F.; Mitjans, M.; Urgatondo, V.; Rocha-Filho, P.A.; Vinardell, M.P. Photoprotective
potential of emulsions formulated with Buriti oil (Mauritia flexuosa) against UV irradiation on
keratinocytes and fibroblasts cell lines. Food Chem. Toxicol. 2010, 48, 70-75.
7 Dhawan, K.; Dhawan, S.; Sharma, A. Passiflora: A review update. J. Ethnopharm. 2004, 94, 1-23.
8 Ripa, F.A.; Haque, M.; Nahar, L.; Islam, Md.M. Antibacterial, cytotoxic and antioxidant activity
of Passiflora edulis Sims. Eur. J. Sci. Res. 2009, 31, 592-598.
9 Facioli, N.L.; Gonçalves, L.A.G. Modificação por via enzimática da composição triglicerídica do
óleo de piqui (Caryocar brasiliense Camb). Quím. Nova 1998, 21, 16-19.
10 Sena, D.M.; Rodrigues, F.F.G.; Freire, P.T.C.; Lima, S.G.; Coutinho, H.D.M.; Carvajal, J.C.L.;
Costa, J.G.M. Physicochemical and spectroscopical investigation of Pequi (Caryocar coriaceum
Wittm.) pulp oil. Grasas Y Aceites 2010, 61, 191-196.
11 Jackson, F.L.; Longenecker, H.E. The fatty acids and glycerides of babassu oil. J. Am. Oil Chem.
Soc. 1944, 21, 73-75.
12 Silva, S.M.; Sampaio, K.A.; Taham, T.; Rocco, S.A.; Ceriani, R.; Meirelles, A.J.A. A
characterization of oil extracted from buriti fruit (Mauritia flexuosa) grown in the Brazilian
Amazon Region. J. Am. Oil Chem. Soc. 2009, 86, 611-616.
13 Kobori, C.N.; Jorge, N. Caracterização dos óleos de algumas sementes de frutas como
aproveitamento de resíduos industriais. Ciência E Agrotecnologia 2005, 29, 1008-1014.
14 ANVISA. Agência Nacional da Vigilância Sanitária, Resolução n°482, 07/23/1999. Available
online: http://www.anvisa.gov.br/legis/resol/482_99.htm (accessed on 07/04/2011).
15 Nyanzi, S.A.; Carstensen, B.; Schwack, W.J. A comparative study of fatty acid profiles of
passiflora seed oils from Uganda. J. Am. Oil Chem. Soc. 2005, 82, 41-44.
16 Quiroga, O.E.; Vigo, M.S. Chemical characteristics of Passiflora caerulea seed oil and residual
seed meal. Molecules 2000, 5, 376-379.
17 Costa, J.G.M.; Brito, S.A.; Nascimentoa, E.M.M.; Botelho, M.A.; Rodrigues, F.F.G.;
Coutinho, H.D.M. Antibacterial properties of pequi pulp oil (Caryocar coriaceum – Wittm.) Int. J.
Food Prop. 2010, in press, doi:10.1080/10942910903207744.
Molecules 2011, 16 5885

18 Azevedo-Meleiro, C.H.; Rodriguez-Amaya, D.B. Confirmation of the identity of the carotenoids

of tropical fruits by HPLC-DAD and HPLC-MS. J. Food Comp. Anal. 2004, 17, 385-396.
19 Rodriguez-Amaya, D.B. Passion Fruits. In Encyclopedia of food sciences and nutrition, 2nd ed.;
Caballero, B., Trugo, L.C., Finglas, P.M., Eds.; Academic Press: Amsterdam, The Netherlands,
2003; Volumes 10, pp. 4368-4373.
20 Rodriguez-Amaya, D.B.; Kimura, M.; Godoy, H.T.; Amaya-Farfan, J. Updated Brazilian database
on food carotenoids: Factors affecting carotenoid composition. J. Food Comp. Anal. 2008, 21,
445-463.
21 Rice-Evans, C.A.; Miller, N.J.; Bolwell, P.G.; Bramley, P.M.; Pridham, J.B. The relative
antioxidant activities of plant derived polyphenolic flavonoids. Free Radic. Res. 1995, 22,
375-383.
22 Teixeira, S.; Mendes, A.; Alves, A.; Santos, L. Simultaneous distillation-extraction of highly
volatile compounds from Cistus ladanifer L. Anal. Chim. Acta 2007, 584, 439-446.
23 Paula, W., Jr.; Rocha, F.H.; Donatti, L.; Fadel-Picheth, C.M.T.; Weffort-Santos, A.M.
Leishmanicidal, antibacterial, and antioxidant activities of Caryocar brasiliense Cambess leaves
hydroethanolic extract. Rev. Bras. Farmacog. 2006, 16, 625-630.
24 Almeida, A.C.; Sobrinho, E.M.; Pinho, L.; Souza, P.N.S.; Martins, E.R.; Duarte, E.R.;
Santos, H.O.; Brandi, I.V.; Cangussu, A.S.; Costa, J.P.R. Acute toxicity of leaf hydroalcoholic
extracts of Lippia sidoides, Myracroduon urundeuva, Stryphnodendron adstringens and of
Caryocar brasiliense administered by intraperitoneal route. Ciência Rural 2010, 40, 200-203.
25 Firestone, D. Official Methods and Recommended Practices of the American Oil Chemists’
Society, 3rd ed.; American Oil Chemists’ Society: Champaign, IL, USA, 1998.
26 AOCS Official Method Ce 2-66. Preparations of Methyl Esters of Fatty Acids. American Oil
Chemists’ Society: Champaign, IL, USA, 1997.
27 Defining, Establishing, and Verifying Reference Intervals in the Clinical Laboratory; Approved
Guideline, In CLSI Document C28-A3, 3rd ed.; Clinical and Laboratory Standards Institute:
Wayne, PA, USA, 2008.
28 Brand-Williams, W.; Cuvelier, M.E.; Berset, C. Use of a free radical method to evaluate
antioxidant activity. Food Sci. Technol. 1995, 28, 25-30.
29 Albano, S.M.; Miguel, M.G. Biological activities of extracts of plants grown in Portugal. Ind.
Crops Prod. 2011, 33, 338-343.
30 Meyer, B.N.; Ferrigni, N.R.; Putnam, L.B.; Jacobsen, L.B.; Nichols, D.E.; McLaughlin, J.L. Brine
shrimp: A convenient general bioassay for active plant constituents. J. Med. Plants Res. 1982, 45,
31-34.

Sample Availability: Samples of the oils are available from the authors.

© 2011 by the authors; licensee MDPI, Basel, Switzerland. This article is an open access article
distributed under the terms and conditions of the Creative Commons Attribution license
(http://creativecommons.org/licenses/by/3.0/).